Investigation of Microbial Community Interactions Between Lake Washington Methanotrophs Using Genome-Scale Metabolic Modeling

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Investigation of Microbial Community Interactions Between Lake Washington Methanotrophs Using Genome-Scale Metabolic Modeling University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Chemical and Biomolecular Engineering -- All Chemical and Biomolecular Engineering, Faculty Papers Department of 2020 Investigation of microbial community interactions between Lake Washington methanotrophs using genome-scale metabolic modeling Mohammad Mazharul Islam Tony Le Follow this and additional works at: https://digitalcommons.unl.edu/chemengall This Article is brought to you for free and open access by the Chemical and Biomolecular Engineering, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Chemical and Biomolecular Engineering -- All Faculty Papers by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Investigation of microbial community interactions between Lake Washington methanotrophs using genome-scale metabolic modeling Mohammad Mazharul Islam1, Tony Le2, Shardhat R. Daggumati1 and Rajib Saha1 1 Department of Chemical and Biomolecular Engineering, University of Nebraska—Lincoln, Lincoln, NE, United States of America 2 Department of Biochemistry, University of Nebraska—Lincoln, Lincoln, NE, United States of America ABSTRACT Background. The role of methane in global warming has become paramount to the environment and the human society, especially in the past few decades. Methane cycling microbial communities play an important role in the global methane cycle, which is why the characterization of these communities is critical to understand and manipulate their behavior. Methanotrophs are a major player in these communities and are able to oxidize methane as their primary carbon source. Results. Lake Washington is a freshwater lake characterized by a methane-oxygen countergradient that contains a methane cycling microbial community. Methanotrophs are a major part of this community involved in assimilating methane from lake water. Two significant methanotrophic species in this community are Methylobacter and Methylomonas. In this work, these methanotrophs are computationally studied via developing highly curated genome-scale metabolic models. Each model was then integrated to form a community model with a multi-level optimization framework. The competitive and mutualistic metabolic interactions among Methylobacter and Methylomonas were also characterized. The community model was next tested under carbon, oxygen, and nitrogen limited conditions in addition to a nutrient-rich Submitted 21 February 2020 condition to observe the systematic shifts in the internal metabolic pathways and Accepted 10 June 2020 Published 30 June 2020 extracellular metabolite exchanges. Each condition showed variations in the methane oxidation pathway, pyruvate metabolism, and the TCA cycle as well as the excretion of Corresponding author Rajib Saha, [email protected] formaldehyde and carbon di-oxide in the community. Finally, the community model was simulated under fixed ratios of these two members to reflect the opposing behavior Academic editor Ludmila Chistoserdova in the two-member synthetic community and in sediment-incubated communities. The community simulations predicted a noticeable switch in intracellular carbon Additional Information and Declarations can be found on metabolism and formaldehyde transfer between community members in sediment- page 19 incubated vs. synthetic condition. DOI 10.7717/peerj.9464 Conclusion. In this work, we attempted to predict the response of a simplified methane cycling microbial community from Lake Washington to varying environments and also Copyright provide an insight into the difference of dynamics in sediment-incubated microcosm 2020 Islam et al. community and synthetic co-cultures. Overall, this study lays the ground for in silico Distributed under systems-level studies of freshwater lake ecosystems, which can drive future efforts of Creative Commons CC-BY 4.0 OPEN ACCESS How to cite this article Islam MM, Le T, Daggumati SR, Saha R. 2020. Investigation of microbial community interactions between Lake Washington methanotrophs using genome-scale metabolic modeling. PeerJ 8:e9464 http://doi.org/10.7717/peerj.9464 understanding, engineering, and modifying these communities for dealing with global warming issues. Subjects Bioinformatics, Computational Biology, Microbiology, Molecular Biology, Freshwater Biology Keywords Methylomonas sp. LW13, Microbial community, Genome-scale metabolic modeling, Methane-cycling community, Lake Washington, Methylobacter tundripaludum 21/22 INTRODUCTION The accelerated rise in worldwide average temperature in recent years is posing a serious threat to the environment, terrestrial ecosystems, human health, economy, and the ultimate survival of the planet earth. About 20 percent of global warming is caused by methane and it is expected to be 86 times more potent than carbon di-oxide in warming the earth over the next two decades (Houghton, Jenkins & Ephraums, 1990; IPCC, 2013). The impacts of the rapid increase in atmospheric methane (Nisbet et al., 2019) are compounded as higher temperatures are associated with an increase in methane production from wetlands and lakes (Yvon-Durocher et al., 2014). Aerobic methanotrophs are mostly gram-negative Proteobacteria that are an integral part of the global carbon cycle (Hanson & Hanson, 1996). They exist in diverse environments such as wetlands, lakes, and the tundra and use the enzyme methane monooxygenase (MMO) to oxidize methane as their sole source of carbon (Hanson & Hanson, 1996). Methane-oxidizing Verrucomicrobia found in geothermal acidic environments are also involved in methane oxidation while solely using carbon dioxide in the Calvin cycle (Carere et al., 2019; Mohammadi et al., 2019; Van Teeseling et al., 2014). The anaerobic methanotrophic archaea are also an important sink of methane that couple methane oxidation with sulfate reduction mediated by sulfate-reducing bacteria (Cui et al., 2015). Thus, methanotrophs act as the primary biological sink for methane (Hanson & Hanson, 1996), consuming up to 90 percent of the methane produced in soil/sediments in addition to the atmospheric methane (Whalen & Reeburgh, 1990; Krause et al., 2017). Methanotrophs have also shown the ability to produce various useful products such as single-cell proteins, biodiesels, biopolymers, and osmo-protectants (Strong, Xie & Clarke, 2015). Lakes act as major sources and sinks for methane and account for 6 to 16 percent of biologically produced methane (IPCC, 2013; Yvon-Durocher et al., 2014). Lake Washington is a freshwater lake characterized by a methane-cycling community where methanotrophs are one of the important functional microbial groups involved in methane oxidation (Yu et al., 2016). It contains a steep vertical counter-gradient of methane and oxygen, and is separated into oxic and anoxic layers where methane production and consumption occur, respectively (Yu et al., 2016). Hence, it can be a model system to better understand methane-cycling communities in lakes and their role in the global methane cycle. Understanding the metabolic interactions in these communities will aid in developing methods to reduce the amount of methane emitted from lakes. A diverse array of microbes exist in the Lake Washington community, where Proteobacteria comprise 33% of the Islam et al. (2020), PeerJ, DOI 10.7717/peerj.9464 2/28 community and includes a major subtype of methanotrophs, Methylococcaceae at a 10% abundance level (Beck et al., 2013). The genus Methylobacter is the most dominant player in Methylococcaceae group at 47.7%, while other major players include Crenothrix, Methylomonas, and Methylomicrobium at 30.0%, 10.8%, and 7.4%, respectively (Beck et al., 2013). Other members include cyanobacteria, bacteriodetes, acidobacteria, and chloroflexi (Beck et al., 2013). Understanding the physiological dynamics and interactions in natural methane-cycling communities, such as the Lake Washington, are crucial to addressing problems concerning methane's role in global warming and leveraging methanotrophs' possible functions in bioremediation and bioproduction. Omics-based techniques and high-throughput sequencing can elucidate important features of the community such as taxonomic information, community composition, and presence of functionally important genes (Temperton & Giovannoni, 2012). However, it is difficult to assign functionality to members of the community and decipher the roles of individual players due to the complexity of the community and the data involved (Zengler, 2009; Zengler & Palsson, 2012). On the other hand, synthetic communities were proven to be efficient models to provide insight into metabolic capabilities and interactions (De Roy et al., 2014). Simple representative community structures can be made to lower complexity, achieve more consistent results, and efficiently elucidate inter-species interactions (De Roy et al., 2014). Certain Lake Washington community members are easier to cultivate in a laboratory setting than other members. For instance, Methylomonas and Methylosinus species previously shown ease of cultivation in the laboratory (Auman et al., 2000). However, Methylobacter species were difficult to isolate and demonstrated poor growth compared to Methylomonas and Methylosinus (Yu et al., 2016). A synthetic community comprising 50 different Lake Washington microbes belonging to 10 methanotrophic, 36 methylotrophic, and 4 non-methanotrophic heterobacteria showed that Methylobacter was outperformed by Methylomonas in the community (Yu et al., 2016).
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